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International Journal of Cancer Jul 2016Amplification of MYCN is the signature genetic aberration of 20-25% of neuroblastoma and a stratifying marker associated with aggressive tumor behavior. The detection of...
Amplification of MYCN is the signature genetic aberration of 20-25% of neuroblastoma and a stratifying marker associated with aggressive tumor behavior. The detection of heterogeneous MYCN amplification (hetMNA) poses a diagnostic dilemma due to the uncertainty of its relevance to tumor behavior. Here, we aimed to shed light on the genomic background which permits hetMNA in neuroblastoma and tied the occurrence to other stratifying markers and disease outcome. We performed SNP analysis using Affymetrix Cytoscan HD arrays on 63 samples including constitutional DNA, tumor, bone marrow and relapse samples of 26 patients with confirmed hetMNA by MYCN-FISH. Tumors of patients ≤18m were mostly aneuploid with numeric chromosomal aberrations (NCAs), presented a prominent MNA subclone and carried none or a few segmental chromosomal aberrations (SCAs). In older patients, tumors were mostly di- or tetraploid, contained a lower number of MNA cells and displayed a multitude of SCAs including concomitant 11q deletions. These patients often suffered disease progression, tumor dissemination and relapse. Restricted to aneuploid tumors, we detected chromosomes with uniparental di- or trisomy (UPD/UPT) in almost every sample. UPD11 was exclusive to tumors of younger patients whereas older patients featured UPD14. In this study, the MNA subclone appears to be constraint by the tumor environment and thus less relevant for tumor behavior in aggressive tumors with a high genomic instability and many segmental aberrations. A more benign tumor background and lower tumor stage may favor an outgrowth of the MNA clone but tumors generally responded better to treatment.
Topics: Adolescent; Aneuploidy; Child; Child, Preschool; Chromosome Aberrations; Chromosome Deletion; Female; Gene Amplification; Genetic Heterogeneity; Humans; In Situ Hybridization, Fluorescence; Infant; Male; N-Myc Proto-Oncogene Protein; Neuroblastoma; Nuclear Proteins; Oncogene Proteins; Polymorphism, Single Nucleotide
PubMed: 26910568
DOI: 10.1002/ijc.30050 -
Surgery Oct 2018Advanced-stage neuroblastoma patients require multiagent chemotherapy. Intratumoral implantation of vincristine-loaded silk gel uses local diffusion to decrease...
BACKGROUND
Advanced-stage neuroblastoma patients require multiagent chemotherapy. Intratumoral implantation of vincristine-loaded silk gel uses local diffusion to decrease orthotopic neuroblastoma tumor growth in mice. We hypothesize that injecting vincristine-loaded silk gel into 8 locations within the tumor, instead of only centrally, decreases the diffusion distance and improves tumor growth suppression.
METHODS
Human neuroblastoma cells, KELLY, were injected into mouse adrenal glands to create orthotopic tumors. After the tumors reached 100 mm by ultrasound, silk gels loaded with 50 µg vincristine were injected centrally or in 8 areas throughout the tumor. Drug-release profile was measured in vitro. Endpoints were tumor size >1,000 mm and histologic examination.
RESULTS
Vincristine-loaded silk gels suppressed tumor growth up to an inflection point (458.7 ± 234.4 mm for central, 514.3 ± 165.8 mm for 8-point injection) before tumor growth accelerated >200 mm over 3 days. The time to inflection point was 6.6 days for central, 13.3 days for 8-point injection (P < .05). Using the sphere volume equation to approximate tumor volume, splitting the volume into 1/8 decreased the diffusion radius by 1/2. Histologic examination confirmed tumor necrosis adjacent to vincristine-loaded silk gel.
CONCLUSION
Injecting vincristine-loaded sustained release silk gel at 8 separate locations halved the diffusion distance and doubled the time for the tumor to reach the growth inflexion point.
Topics: Animals; Antineoplastic Agents; Biocompatible Materials; Cell Line, Tumor; Diffusion; Drug Delivery Systems; Gels; Humans; Injections, Intralesional; Mice; Neuroblastoma; Silk; Tissue Extracts; Tumor Burden; Vincristine; Xenograft Model Antitumor Assays
PubMed: 30061039
DOI: 10.1016/j.surg.2018.06.017 -
Practical Laboratory Medicine Apr 2017Metastatic disease is a major challenge for cancer cure, haematogenous spread and subsequent growth of tumour cells at distant sites being the cause of most cancer...
Metastatic disease is a major challenge for cancer cure, haematogenous spread and subsequent growth of tumour cells at distant sites being the cause of most cancer deaths. Molecular characterization and detection of the tumour cells responsible for haematogenous spread may increase understanding of the biology of metastasis, help improve patient management and allow evaluation of novel treatments to prevent and eradicate this disease. The bone marrow is a common site to which tumour cells metastasize, from which they may re-circulate to other organs with a favourable microenvironment for growth. The detection of tumour cells in blood suggests one route for metastasis, and provides an accessible, minimally invasive liquid sample through which it may be possible to monitor and detect minimal disease and early signs of metastasis. Significant improvements in the sensitivity and specificity of tumour cell detection have been made, such that it is now possible to unambiguously detect a single tumour cell in over 10 million normal cells. However, the clinical impact of such low level disease and how to interpret the natural variation that can arise from sequential sampling of bone marrow aspirates and blood is currently largely unknown. This commentary will focus on the technical advancements and application of reverse transcriptase polymerase chain reaction to detect cancer mRNAs in bone marrow and blood, and discuss the potential clinical impact of this test in neuroblastoma.
PubMed: 28856217
DOI: 10.1016/j.plabm.2016.04.003 -
European Archives of... Dec 2016Ion beam therapy has enabled us to treat formerly untreatable malignant tumors. The aim of the present study was to investigate the long-term follow-up course of...
Ion beam therapy has enabled us to treat formerly untreatable malignant tumors. The aim of the present study was to investigate the long-term follow-up course of patients with head and neck cancers who received ion beam therapy. The subjects were 8 patients (3 men and 5 women aged 43-78 years) with head and neck cancers who visited our department from 2006 to 2015 and received ion beam therapy. Six patients received carbon ion beam therapy, and the other two patients received proton beam therapy. The medical records of the patients were retrospectively analyzed. The primary site was the nasal and paranasal sinuses in six cases, nasopharynx in one case, and external auditory canal in one case. The histological type was olfactory neuroblastoma, malignant melanoma, and adenoid cystic carcinoma in two cases each, and chondrosarcoma and squamous cell carcinoma in one case each. The exposure dose ranged from 64 to 70.4 GyE. The average follow-up period was 42.0 months. Early adverse events were generally mild, and complete therapeutic response was obtained in all cases. However, five patients developed severe late complications including craniospinal dissemination, osteoradionecrosis of the maxilla and skull base, brain necrosis, and loss of eyesight. Three patients died of distant metastasis, local recurrence and/or brain necrosis within 2 years, and four patients have been surviving with distant metastasis or severe late complications. Ion beam therapy exhibits outstanding antitumor effects, but the severe late complications of the therapy must also be recognized.
Topics: Adult; Aged; Carcinoma, Adenoid Cystic; Carcinoma, Squamous Cell; Esthesioneuroblastoma, Olfactory; Female; Head and Neck Neoplasms; Heavy Ion Radiotherapy; Humans; Male; Melanoma; Middle Aged; Proton Therapy; Radiotherapy Dosage; Retrospective Studies; Treatment Outcome
PubMed: 27168403
DOI: 10.1007/s00405-016-4086-2 -
Journal of Virology Jun 2017Although a varicella-zoster virus (VZV) vaccine has been used for many years, the neuropathy caused by VZV infection is still a major health concern. Open reading frame...
Although a varicella-zoster virus (VZV) vaccine has been used for many years, the neuropathy caused by VZV infection is still a major health concern. Open reading frame 7 (ORF7) of VZV has been recognized as a neurotropic gene , but its neurovirulent role remains unclear. In the present study, we investigated the effect of ORF7 deletion on VZV replication cycle at virus entry, genome replication, gene expression, capsid assembly and cytoplasmic envelopment, and transcellular transmission in differentiated neural progenitor cells (dNPCs) and neuroblastoma SH-SY5Y (dSY5Y) cells. Our results demonstrate that the ORF7 protein is a component of the tegument layer of VZV virions. Deleting ORF7 did not affect viral entry, viral genome replication, or the expression of typical viral genes but clearly impacted cytoplasmic envelopment of VZV capsids, resulting in a dramatic increase of envelope-defective particles and a decrease in intact virions. The defect was more severe in differentiated neuronal cells of dNPCs and dSY5Y. ORF7 deletion also impaired transmission of ORF7-deficient virus among the neuronal cells. These results indicate that ORF7 is required for cytoplasmic envelopment of VZV capsids, virus transmission among neuronal cells, and probably the neuropathy induced by VZV infection. The neurological damage caused by varicella-zoster virus (VZV) reactivation is commonly manifested as clinical problems. Thus, identifying viral neurovirulent genes and characterizing their functions are important for relieving VZV related neurological complications. ORF7 has been previously identified as a potential neurotropic gene, but its involvement in VZV replication is unclear. In this study, we found that ORF7 is required for VZV cytoplasmic envelopment in differentiated neuronal cells, and the envelopment deficiency caused by ORF7 deletion results in poor dissemination of VZV among neuronal cells. These findings imply that ORF7 plays a role in neuropathy, highlighting a potential strategy to develop a neurovirulence-attenuated vaccine against chickenpox and herpes zoster and providing a new target for intervention of neuropathy induced by VZV.
Topics: Capsid; Cell Differentiation; Cell Line; Cytoplasm; Gene Deletion; Genome, Viral; Herpes Zoster; Herpesvirus 3, Human; Humans; Neuroblastoma; Neurons; Viral Envelope Proteins; Viral Proteins; Virion; Virus Internalization; Virus Replication
PubMed: 28356523
DOI: 10.1128/JVI.00127-17 -
Polish Journal of Pathology : Official... Jun 2016Olfactory neuroblastoma (ONB) is a rare neoplasm of the sinonasal area with neuroendocrine differentiation. ISL-1, TTF-1 and PAX5 are transcription factors that are...
Olfactory neuroblastoma (ONB) is a rare neoplasm of the sinonasal area with neuroendocrine differentiation. ISL-1, TTF-1 and PAX5 are transcription factors that are frequently upregulated in tumors showing neuroendocrine differentiation. The aim of our study was to evaluate these markers in a group of ONBs. We included 11 ONBs from 4 large university hospitals. Immunohistochemical expression of TTF-1, PAX5 and ISL-1 was evaluated. TTF-1, ISL-1 and PAX5 were expressed in 3/11 cases (27.27%, h-score: 3-45), 7/11 cases (63.64%, h-score: 23-200), and in 3/11 cases (27.77%, h-score 3-85), respectively. The patient with the strongest PAX5 reactivity exhibited an aggressive clinical course with rapid dissemination to the spine and death shortly after the diagnosis. No significant correlation in the expression of PAX5 and TTF-1 ( = 0.43; p = 0.18) was observed. ISL-1 is widely expressed in tumors with neuroendocrine differentiation and therefore of limited value in their differential diagnosis. TTF-1 positivity does not exclude the diagnosis of primary ONB, although usually only a small percentage of cells are positive. PAX5 expression is infrequent (27.27%) in ONB; however, if present it can be associated with a very aggressive clinical course.
Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; DNA-Binding Proteins; Esthesioneuroblastoma, Olfactory; Female; Humans; Immunohistochemistry; LIM-Homeodomain Proteins; Male; Middle Aged; Nasal Cavity; Nose Neoplasms; PAX5 Transcription Factor; Transcription Factors; Young Adult
PubMed: 27543867
DOI: 10.5114/pjp.2016.61448 -
Integrin α4 Enhances Metastasis and May Be Associated with Poor Prognosis in MYCN-low Neuroblastoma.PloS One 2015High-risk neuroblastoma is associated with an overall survival rate of 30-50%. Neuroblastoma-expressed cell adhesion receptors of the integrin family impact cell...
High-risk neuroblastoma is associated with an overall survival rate of 30-50%. Neuroblastoma-expressed cell adhesion receptors of the integrin family impact cell adhesion, migration, proliferation and survival. Integrin α4 is essential for neural crest cell motility during development, is highly expressed on leukocytes, and is critical for transendothelial migration. Thus, cancer cells that express this receptor may exhibit increased metastatic potential. We show that α4 expression in human and murine neuroblastoma cell lines selectively enhances in vitro interaction with the alternatively spliced connecting segment 1 of fibronectin, as well as vascular cell adhesion molecule-1 and increases migration. Integrin α4 expression enhanced experimental metastasis in a syngeneic tumor model, reconstituting a pattern of organ involvement similar to that seen in patients. Accordingly, antagonism of integrin α4 blocked metastasis, suggesting adhesive function of the integrin is required. However, adhesive function was not sufficient, as mutants of integrin α4 that conserved the matrix-adhesive and promigratory function in vitro were compromised in their metastatic capacity in vivo. Clinically, integrin α4 is more frequently expressed in non-MYNC amplified tumors, and is selectively associated with poor prognosis in this subset of disease. These results reveal an unexpected role for integrin α4 in neuroblastoma dissemination and identify α4 as a potential prognostic indicator and therapeutic target.
Topics: Animals; Cell Adhesion; Cell Line, Tumor; Cell Movement; Gene Expression Regulation, Neoplastic; Humans; Integrin alpha4; Intercellular Signaling Peptides and Proteins; Liver Neoplasms; Lymphatic Metastasis; Mice; N-Myc Proto-Oncogene Protein; Neoplasm Transplantation; Nervous System Neoplasms; Neuroblastoma; Nuclear Proteins; Oncogene Proteins; Peptides; Prognosis; Signal Transduction; Survival Analysis; Vascular Cell Adhesion Molecule-1
PubMed: 25973900
DOI: 10.1371/journal.pone.0120815 -
Journal of Virological Methods Sep 2017Maintaining a healthy, continuous immortalized cell line is essential for rabies laboratories that perform virus isolation assays and test for the presence of viral...
Maintaining a healthy, continuous immortalized cell line is essential for rabies laboratories that perform virus isolation assays and test for the presence of viral neutralizing antibodies. Individuals who routinely work with rabies virus, such as rabies laboratory employees, or those who may have a high potential for exposure to rabies virus, including veterinarians, should be tested for the presence of anti-rabies viral neutralizing antibodies (VNA) every 6-24 months, depending on potential exposure level. The gold standard for serum neutralization assays require the use of live rabies virus and cells that are sensitive to rabies virus infection. Additionally, virus isolation assays are routinely performed in rabies laboratories as a back-up for the direct fluorescent antibody test (dFAT). Currently there are no guidelines or publications recommending the use of low, intermediate, or high passage cell lines in rabies assays. In this study, we compared the sensitivity of intermediate, high, and extremely high passaged neuroblastomas to rabies virus using virus isolation, serum neutralization, and real time RT-PCR techniques. Additionally, cells were examined microscopically to determine changes in morphology and dissemination of rabies virus antigen between intermediate, high, and extremely high passage cells. No significant difference was found between cell passage numbers and viral susceptibility between intermediate and high passaged cells. However, extremely high passaged cells (≥1200 passages) were less susceptible to viral infection and/or produced less virus following inoculation. As a result, rabies laboratories that use viral isolation and serum neutralization assays should regularly assess cell susceptibility to ensure the integrity and repeatability of the test.
Topics: Cell Line, Tumor; Humans; Neurons; Neutralization Tests; Rabies virus; Serial Passage; Tissue Culture Techniques; Viral Tropism; Virus Cultivation
PubMed: 28506631
DOI: 10.1016/j.jviromet.2017.05.005 -
International Journal of Environmental... Feb 2021Neuroblastoma is the most common extra-cranial solid tumor in infants and young children, and accounts for approximately 8-10% of all childhood cancers. The...
Neuroblastoma is the most common extra-cranial solid tumor in infants and young children, and accounts for approximately 8-10% of all childhood cancers. The International Neuroblastoma Staging System (The International Neuroblastoma Risk Group Staging System (INRGSS)) is based on the age of patient and preoperative imaging, with attention paid to whether the primary tumor is affected by one or more of specific Image-Defined Risk Factors (IDRFs). Patients are classified into the following groups: locoregional L1 and L2 (absent or present IDRFs respectively), M stage (a disseminated form of neuroblastoma) and Ms (the stage present in children younger than 18 months of age with the disease spread to the bone marrow and/or liver, and/or skin). This publication is aimed to present an unexpected complication associated with an accidental ligation of the celiac trunk during resection of a neuroblastoma tumor in a 2.5-year-old boy after initial chemotherapy, initially with vascular IDRFs, stage L2. The consequences of this complication were pancreatic and spleen ischemia and necrosis, and ischemia and perforation of the common bile duct, gallbladder, stomach, and duodenum. Despite detailed diagnostic imaging (computed tomography, magnetic resonance), the presence of vascular IDRFs may result in an unexpected complication in the surgical treatment of neuroblastoma in children.
Topics: Child; Child, Preschool; Family; Humans; Infant; Magnetic Resonance Imaging; Male; Neoplasm Staging; Neuroblastoma; Risk Factors; Tomography, X-Ray Computed
PubMed: 33672809
DOI: 10.3390/ijerph18041841 -
Cirugia Pediatrica : Organo Oficial de... Aug 2018The aim of the paper is to describe the experience in our center with the use of minimally invasive surgery (MIS) of neural tumors in childhood.
PURPOUSE
The aim of the paper is to describe the experience in our center with the use of minimally invasive surgery (MIS) of neural tumors in childhood.
METHODS
Descriptive and retrospective study of patients diagnosed with neural neoplasia (neuroblastoma and neuroganglioma) on whom MIS technique surgery has been performed between October 2012 and December 2017. The inclusion criteria were patients with a neural tumor diagnosis who, at the time of the intervention, did not have imaging-defined risk factors (IDRFs). Patients with a different diagnosis than neural tumor or with IDRFs were excluded from the study.
RESULTS
The study comprises 19 cases (6 female and 13 male) with a median age of 47 months. According to the International Neuroblastoma Risk Group Staging System (INRGSS) classification, nine cases were in L1 stage, six in L2, two in M and two in MS. Laparoscopy was used in 14 patients (12 adrenal and 2 abdominal tumors) and thoracoscopy was used in the other 5. In 4 of the 19 cases (21%), conversion to open surgery was needed due to fibrosis in 2 cases and vascular structures entrapment in another 2 (3 in laparoscopy and 1 in thoracoscopy). There were no surgical complications, achieving complete resection in all cases. Three cases showed postsurgical adverse effects grade I and II, according to Clavien-Dindo classification. After a median of 27 months of follow up, two patients showed disease progression without local recurrence.
CONCLUSIONS
In conclusion, MIS are useful techniques in the surgical exeresis of non-disseminated neural tumors without IDRFs.
Topics: Abdominal Neoplasms; Adolescent; Adrenal Gland Neoplasms; Child; Child, Preschool; Disease Progression; Female; Follow-Up Studies; Ganglioneuroma; Humans; Infant; Laparoscopy; Male; Minimally Invasive Surgical Procedures; Neuroblastoma; Retrospective Studies; Risk Factors; Thoracoscopy
PubMed: 30260107
DOI: No ID Found